Critical questions must be addressed to evaluate the potential of metabolomics for studying free-living wildlife. First, can metabolomics identify stress-induced phenotypes in animals experiencing a highly variable environment or must animals be stabilized in a controlled laboratory prior to sampling? Second, is knowledge of species and phenotype (gender and age) required to interpret metabolomics data? To address these questions, we characterized the metabolic variability of the mussel and determined if inherent variability masked the metabolic response to an environmental stressor, hypoxia. Specifically, we compared metabolic fingerprints of adductor muscle and mantle from four groups of Mytilus galloprovincialis: animals sampled directly from the field with and without hypoxia and those stabilized in a laboratory for 60 h, also with and without hypoxia. Contrary to expectation, laboratory stabilization increased metabolic variability in adductor muscle, thereby completely masking the response to hypoxia. The principal source of metabolic variability in mantle was shown to be gender-based, highlighting the importance of phenotypic anchoring of samples to known life history traits. We conclude that direct field sampling is recommended for environmental metabolomics since it minimizes metabolic variability and enables stress-induced phenotypic changes to be observed. Furthermore, we recommend that species and phenotype of the study organism must be known for meaningful interpretation of metabolomics data.

Tilapia lake virus (TiLV), a negative sense RNA virus with a 10 segment genome, is an emerging threat to tilapia aquaculture worldwide, with outbreaks causing over 90% mortality reported on several continents since 2014. Following a severe tilapia mortality event in July 2017, we confirmed the presence of TiLV in Bangladesh and obtained the near-complete genome of this isolate, BD-2017. Phylogenetic analysis of the concatenated 10 segment coding regions placed BD-2017 in a clade with the two isolates from Thailand, separate from the Israeli and South American isolates. However, phylogenetic analysis of individual segments gave conflicting results, sometimes clustering BD-2017 with one of the Israeli isolates, and splitting pairs of isolates from the same region. By comparing patterns of topological difference among segments of quartets of isolates, we showed that TiLV likely has a history of reassortment. Segments 5 and 6, in particular, appear to have undergone a relatively recent reassortment event involving Ecuador isolate EC-2012 and Israel isolate Til-4-2011. The phylogeny of TiLV isolates therefore depends on the segment sequenced. Our findings illustrate the need to exercise caution when using phylogenetic analysis to infer geographic origin and track the movement of TiLV, and we recommend using whole genomes wherever possible.

ABSTRACT:We assessed seasonal histological changes as markers of health status in mussels Mytilus spp. sampled from Southampton Water, Hampshire, UK and the River Exe, Devon, UK between November 2004 and October 2005. A total of 29 health parameters related to pathogens, inflammatory and non-specific pathologies, and reproductive and physiological condition were recorded monthly from individual mussels collected from these 2 sites. We then assessed the diffential prevalence of these health parameters according to species. M. edulis, M. galloprovincialis and their hybrids were identified using the Glu-5' gene and the ME15 and ME16 primer sets that distinguish alleles specific to M. edulis (180 bp), M. galloprovincialis (126 bp) and hybrids (180 bp/126 bp). Although no overall annual differences were observed between species with respect to median levels of adipogranular (ADG) tissue and reproductive status, specific differences in reproductive status were observed within individual months. During these months (August to October), M. edulis exhibited a relatively lower reproductive status compared to M. galloprovincialis and hybrids. With respect to all remaining health parameters (pathogens, inflammatory and non-specific pathology), principal components analysis revealed no overall differences between species throughout the year. However, greater differences were observed between species during the autumn and winter than during the spring and summer, thus indicating that species differences may be exacerbated by season. This study highlights how species can affect the accurate interpretation of histopathology data collected during biological effects monitoring programmes. Whether species can also affect the biomarker response of Mytilus mussels to contaminated environments remains to be shown. The results are discussed in the context of biological effects monitoring utilising mussels.

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